How to amplify 1kb PCR product
Dr. Duncan Clark
duncan at genesys.demon.co.uk
Mon Mar 2 12:38:14 EST 1998
In article <34FAD3B3.4B82 at public1.guangzhou.gd.cn>, gzzhjr <gzzhjr at PUBLI
>The length of DNA fragment I want to amplified by PCR is about 1kb,but I
>have no experience with this,could someone tell me the appropriate
>temperature and time in denature,annealing and expansion process?
>Thanks for any information.
For Taq polymerase use the usual extension time allowed is 1kb per
minute used at 72C. Denaturation on a block machine is usually 94C for
30 sec, annealing is for 30 secs at a temperature decided by your oligo
design. Use 2.5u Taq per 100ul PCR.
The problem with being on the cutting edge is that you occasionally get
sliced from time to time....
More information about the Methods